The concentration of pineapple juice is needed to retain the bromelain activity and to standardize the composition and proteolytic activity. Thus, this work aimed to obtain a pure bromelain extract from the Ananas comosus L. Merril juice by membrane separation process. A 2 2 experimental planning was used to study the influence of pH and transmembrane pressure on the activity recovery by micro-filtration using a plain membrane. In second step, this enzyme was purified by the ultra-filtration using a 10 kDa millipore kit. The best operation condition to bromelain concentration using the plain membrane was at pH 7.5 and transmembrane pressure of 0.05 bar, while 85% of bromelain activity was recovered. Ultra-filtration retained 100% of proteolytic activity and concentrated in 10 fold the bromelain extract. SDS-PAGE electrophoresis showed that the ultra-filtrated had high purity and the bromelain from A. comosus pulp had a molecular weight of 24.5 kDa.
Infections caused by Streptococcus pneumoniae are one of the main causes of death around the world. In order to address this problem, investigations are being made into the development of a protein-based vaccine. The aims of this study were to clone and express ClpP, a protein from S. pneumoniae serotype 14 in Escherichia coli, to optimize protein expression by using experimental design and to study plasmid segregation in the system. ClpP was cloned into the pET28b vector and expressed in E. coli BL21 Star (DE3). Protein expression was optimized by using central composite design, varying the inducer (IPTG) and kanamycin concentration, with a subsequent analysis being made of the concentration of heterologous protein, cell growth and the fraction of plasmid-bearing cells. In all the experiments, approximately the same concentration of ClpP was expressed in its soluble form, with a mean of 240.4mg/L at the center point. Neither the IPTG concentration nor the kanamycin concentration was found to have any statistically significant influence on protein expression. Also, higher IPTG concentrations were found to have a negative effect on cell growth and plasmid stability. Plasmid segregation was identified in the system under all the concentrations studied. Using statistical analysis, it was possible to ascertain that the procedures for determining plasmid stability (serial dilution and colony counting) were reproducible. It was concluded that the inducer concentration could be reduced tenfold and the antibiotic eliminated from the system without significantly affecting expression levels and with the positive effect of reducing costs.
Adsorption equilibrium is a fundamental concept in the adsorption science and relates the equilibrium between the quantity of the adsorbed material and its concentration in the bulk phase. Several models have been proposed for prediction of adsorption equilibrium and all models depend on parameters whose values must be estimated from available experimental data. Although linear parameter estimation procedures can be used for model fitting, through transformation of available experimental data and model parameters, non-linear parameter estimation procedures lead to more reliable results and allow for direct comparison of results obtained with different adsorption equilibrium models. The main objective of this work is to present and compare different non-linear procedures for parameter estimation of adsorption equilibrium models, based on theoretical arguments and also on the numerical estimation of adsorption equilibrium parameters, using available experimental data for adsorption of methylene blue onto activated carbon. The results obtained indicate that the best parameter estimation procedure is the one that relies on available equilibrium concentrations in the bulk phase as a function of the fluid volume, adsorbent mass and initial concentrations in the bulk phase, without transformation of measured experimental values and model parameters. Besides, it is shown that parameter estimates should be obtained through proper minimization of weighted least-squares objective function, in accordance with maximum likelihood procedures.
Neste trabalho, estudou-se o comportamento hidrodinâmico de uma coluna de adsorção em leito expandido com o objetivo de obter as condições ótimas para recuperação de amilases do malte de milho. Usou-se a resina de troca iônica Amberlite IRA 410, o tampão fosfato pH 6 em 0,07 M, sem e com 2 % do malte de milho, e água destilada como líquidos fluidizantes, nas temperaturas 295,15 ± 1 K e 301,15 ± 1 K. A equação de Richardson-Zack se ajustou perfeitamente à descrição do comportamento da fluidização dos adsorventes em leito expandido, e as velocidades terminais experimentais apresentaram um erro relativo abaixo dos 15% com relação à velocidade teórica, o que mostra que essas equações são válidas e podem também ser aplicadas em adsorção em leito expandido. O estudo de distribuição do tempo de residência mostrou que houve um aumento da Daxial e do Pe com a elevação da temperatura, da altura do leito e da porosidade. O N foi constante e igual a 2,14, sendo a condição ótima de recuperação de biomoléculas do malte de milho aquela que possuiu menor porosidade do leito fluidizado, por apresentar maior viabilidade econômica.
The proper determination of experimental errors in bioprocesses can be very important because experimental errors can exert a major impact on the analysis of experimental results. Despite this, the effect of experimental errors on the analysis of bioprocess data has been largely overlooked in the literature. For this reason, we performed detailed statistical analyses of experimental errors obtained during the production of lactobionic acid and sorbitol in a system utilizing as catalyst the GFOR (glucose-fructose oxidoreductase) enzyme from permeabilized cells of the bacteria Zymomonas mobilis. The magnitude of the experimental errors thus obtained were then correlated with the process operation conditions and with the composition of the culture media used for bacterial growth. It is shown that experimental errors can depend very significantly on the operation conditions and affect the interpretation of available experimental data. More specifically, in this study, experimental errors depended on the nutritional supplements added to the cultivation medium, the inoculation process, and the reaction time, which may be of fundamental importance for actual process development. The results obtained also indicate, for the first time, that GFOR activity can be affected by the composition of the medium in which cells are cultivated.
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